ATCC medium: 1653 Revised magnetic Spirillum growth medium (MSGM)
Distilled water...........................1.0 L
Wolfe's Vitamin Solution (see below)......10.0 ml
Wolfe's Mineral Solution (see below)......5.0 ml
0.01 M Ferric Quinate (see below).........2.0 ml
0.1% Resazurin............................0.45 ml
KH2PO4 ....................................0.68 g
NaNO3 .....................................0.12 g
Ascorbic acid.............................0.035 g
Tartaric acid.............................0.37 g
Succinic acid.............................0.37 g
Sodium acetate............................0.05 g
Agar (for semi-solid medium)..............1.3 g
Add components in the order given with stirring.
Adjust to pH 6.75 with NaOH. Liquid Medium: Autoclave medium at 121C for 15 minutes. Aseptically fill screw-capped containers to full capacity with sterile medium. Inoculate heavily leaving no headspace of air, and screw down closures tightly.
Semi-Solid Medium: Dispense 12 ml per 16 X 150 mm screw-cap tube.
Autoclave at 121C for 15 minutes.
0.01 M Ferric Quinate:
FeCl3 .....................................0.27 g
Quinic Acid (Sigma Q-0500)................0.19 g
Distilled water...........................100.0 ml
Dissolve and autoclave at 121C for 15 minutes.
Wolfe's Vitamin Solution:
Available from ATCC as a sterile ready-to-use liquid (Vitamin
Supplement, catalog no.MD-VS).
Biotin....................................2.0 mg
Folic acid................................2.0 mg
Pyridoxine hydrochloride..................10.0 mg
Thiamine . HCl............................5.0 mg
Riboflavin................................5.0 mg
Nicotinic acid............................5.0 mg
Calcium D-(+)-pantothenate................5.0 mg
Vitamin B12...............................0.1 mg
p-Aminobenzoic acid.......................5.0 mg
Thioctic acid.............................5.0 mg
Distilled water...........................1.0 L
Wolfe's Mineral Solution:
Available from ATCC as a sterile ready-to-use liquid (Trace Mineral
Supplement, catalog no. MD-TMS.
) Nitrilotriacetic acid.....................1.5 g
MgSO4 . 7H2O ..............................3.0 g
MnSO4 . H2O ...............................0.5 g
NaCl......................................1.0 g
FeSO4 . 7H2O ..............................0.1 g
CoCl2 . 6H2O ..............................0.1 g
CaCl2 .....................................0.1 g
ZnSO4 . 7H2O ..............................0.1 g
CuSO4 . 5H2O ..............................0.01 g
AlK(SO4)2 . 12H2O..........................0.01 g
H3BO3 .....................................0.01 g
Na2MoO4 . 2H2O.............................0.01 g
Distilled water...........................1.0 L
Add nitrilotriacetic acid to approximately 500 ml of water and adjust to pH 6.5 with KOH to dissolve the compound. Bring volume to 1.0 L with remaining water and add remaining compounds one at a time.